This invention relates to compounds that inhibit farnesylation of mutant ras gene products through inhibition of the enzyme farnesyl-protein transferase (FPTase). The invention also relates to methods of manufacturing the compounds, pharmaceutical compositions and methods of treating diseases, especially cancer, which are mediated through farnesylation of ras.
Cancer is believed to involve alteration in expression or function of genes controlling cell growth and differentiation. Whilst not wishing to be bound by theoretical considerations the following text sets out the scientific background to ras in cancer. Ras genes are frequently mutated in tumours. Ras genes encode guanosine triphosphate (GTP) binding proteins which are believed to be involved in signal transduction, proliferation and malignant transformation. H-, K- and N-ras genes have been identified as mutant forms of ras (Barbacid M. Ann. Rev. Biochem. 1987, 56: 779-827). Post translational modification of ras protein is required for biological activity. Farnesylation of ras catalysed by FPTase is believed to be an essential step in ras processing. It occurs by transfer of the farnesyl group of farnesyl pyrophosphate (FPP) to a cysteine at the C-terminal tetrapeptide of ras in a structural motif called the CAAX box. After further post-translational modifications. including proteolytic cleavage at the cysteine residue of the CAAX box and methylation of the cysteine carboxyl, ras is able to attach to the cell membrane for relay of growth signals to the cell interior. In normal cells activated ras is believed to act in conjunction with growth factors to stimulate cell growth. In tumour cells it is believed that mutations in ras cause it to stimulate cell division even in the absence of growth factors (Travis J. Science 1993, 260: 1877-1878), possibly through being permanently in GTP activated form rather than cycled back to GDP inactivated form. Inhibition of farnesylation of mutant ras gene products will stop or reduce activation.
One class of known inhibitors of farnesyl transferase is based on farnesyl pyrophosphate analogues: see for example European patent application EP 534546 from Merck, inhibitors of farnesyl transferase based on mimicry of the CAAX box have been reported. Reiss (1990) in Cell 62, 81-8 disclosed tetrapeptides such as CVIM (Cys-Val-Ile-Met). James (1993) in Science 260, 1937-1942 disclosed benzodiazepine based peptidomimetic compounds. After earliest priority date of the present invention Lerner (1995) in J. Biol. Chem. 270, 26802 and Eisai in International Patent Application WO 95/25086 disclosed further peptidomimetic compounds based on Cys as the first residue. Also after the earliest priority date of the present invention Bristol-Myers Squibb in European Patent Application EP 696593 disclosed for the first time farnesyl transferase inhibitors having a 4-sulfanylpyrrolidine residue in the first position.
According to one aspect of the present invention there is provided a pharmaceutical composition comprising an inhibitor of ras farnesylation of Formula I 
wherein:
R1 is selected from H; xe2x80x94C1-4alkyl; xe2x80x94C1-3alkylene-Ph optionally mono or di-substituted on Ph with substituents selected from C1-4alkyl, halogen, OH, C1-4alkoxy, C1-4alkanoyl, C1-4alkanoyloxy, amino, C1-4alkylamino, di(C1-4alkyl)amino, C1-4alkanoylamino, nitro, cyano, carboxy, carbamoyl, C1-4alkoxycarbonyl, thiol, C1-4alkylsulfanyl, C1-4alkylsulfinyl, C1-4alkylsulfonyl and sulfonamido; xe2x80x94COxe2x80x94C1-4alkyl; xe2x80x94COxe2x80x94Oxe2x80x94C1-4alkyl; xe2x80x94COxe2x80x94Oxe2x80x94C2-4alkenyl; xe2x80x94COxe2x80x94Oxe2x80x94(CH2)nPh optionally substituted on Ph as defined for substitution on Ph in R1=xe2x80x94C1-3alkylene-Ph above and n=0-4; xe2x80x94C1-4alkylene-CONR4R5 where R4 and R5 are independently selected from H and C1-4alkyl; and xe2x80x94C1-4alkylene-COOR6 where R6 is selected from H, C1-4alkyl;
R2 is selected from H; xe2x80x94C1-4alkyl; xe2x80x94C1-3alkylene-Ph optionally substituted on Ph as defined for substitution on Ph in R1=xe2x80x94C1-3alkylene-Ph above; xe2x80x94COC1-4alkyl; and xe2x80x94COOC1-4alkyl;
R3 is selected from H; OH; CN; CF3; NO2; xe2x80x94C1-4 alkyl; xe2x80x94C1-4alkylene-R7 where R7 is selected from phenyl, naphthyl, a 5-10 membered monocyclic or bicyclic heteroaryl ring containing up to 5 heteroatoms selected from O, N and S and any aryl ring in R7 is optionally substituted as defined for substitution on the Ph group in R1=xe2x80x94C1-3alkylene-Ph above; R7; C2-4alkenyl; halogen; xe2x80x94(CH2)nCOOR8 where n=0-3 and R8 represents H, C1-4alkyl, or C2-4alkenyl; xe2x80x94CONR9R10 where R9 and R10 independently represent H. C1-4alkyl, C2-4alkenyl, xe2x80x94Oxe2x80x94C1-4alkyl, xe2x80x94Oxe2x80x94C2-4alkenyl, xe2x80x94C1-3alkylenePh optionally substituted as defined for this group for R1 above: xe2x80x94CON(R11)OR12 where R11 and R12 independently represent H, C1-4alkyl and C2-4alkenyl; a group of Formula II, xe2x80x94CONR13xe2x80x94CHR14xe2x80x94COOR17, where R13 is H or C1-4alkyl. R17 is H or C1-6alkyl. R14 is selected from the side chain of a lipophilic amino acid. carbamoylC1-4alkyl, N-(monoC1-4alkyl)carbamoylC1-4alkyl and N-(diC1-4alkyl)carbamoylC1-4alkyl, the group of Formula II having L or D confieuration at the chiral alpha carbon in the corresponding free amino acid: a lactone of formula 
C1-4alkyl monosubstituted on carbon with xe2x95x90Nxe2x80x94OH; a group of Formula xe2x80x94Xxe2x80x94R15 where X is selected from O, CO, CH2, S, SO, SO2 and R15 is selected from C1-6alkyl, phenyl, naphthyl, a 5-10 membered monocyclic or bicyclic heteroaryl ring containing up to 5 heteroatoms selected from O, N and S and any aryl ring in R15 is optionally substituted as defined for the Ph group in R1=xe2x80x94C1-3alkylene-Ph; p is 0-3, in which R3 values can be the same or different: L is a linking moiety selected from the following groups written from left to right in Formula I: xe2x80x94COxe2x80x94NR16xe2x80x94 where R16 is selected from H, C1-4alkyl, C1-4alkylene-Z, xe2x80x94COxe2x80x94C1-4alkylene-Z, xe2x80x94COxe2x80x94C1-6alkyl, xe2x80x94COZ, Z and Z is selected from xe2x80x94Oxe2x80x94C1-4alkyl, phenyl, naphthyl, a 5-10 membered monocyclic or bicyclic heteroaryl ring containing up to 5 heteroatoms selected from O, N and S and any aryl ring in R16 is optionally substituted as defined for the Ph group in R1=xe2x80x94C1-3alkylene-Ph; xe2x80x94CH2-NR18xe2x80x94 where R18 represents any value defined for R16; xe2x80x94CH2Sxe2x80x94; xe2x80x94CH2Oxe2x80x94, xe2x80x94CH2CHR19xe2x80x94 where R19 represents any value defined for R16; xe2x80x94CHxe2x95x90CR20xe2x80x94 where R20 represents any value defined for R16; xe2x80x94CH2NR21xe2x80x94Txe2x80x94 where R21 represents any value defined for R16, T represents xe2x80x94(CH2)nxe2x80x94 where n is 1-4 and T is optionally monosubstituted with R22 where R22 represents any value for R16 other than H; xe2x80x94CHNR2xe2x80x94SO2xe2x80x94 where R23 represents any value defined for R16; xe2x80x94CH2NR24xe2x80x94COxe2x80x94Txe2x80x94 where R24 represents any value defined for R16, T represents xe2x80x94(CH2)nxe2x80x94 where n is 0-4 and T is optionally monosubstituted with R29 where R29 represents any value for R16 other than H; xe2x80x94COxe2x80x94NR25xe2x80x94Txe2x80x94 where R25 represents any value defined for R16, T represents xe2x80x94(CH2)nxe2x80x94 where n is 1-4 and T is optionally monosubstituted with R26 where R26 represents any value for R16 other than H, xe2x80x94CH2Sxe2x80x94Txe2x80x94 where T represents xe2x80x94(CH2)nxe2x80x94 where n is 1-4 and T is optionally monosubstituted with R27 where R27 represents any value for R16 other than H; xe2x80x94CH2Oxe2x80x94Txe2x80x94 where T represents xe2x80x94(CH2)nxe2x80x94 where n is 1-4 and T is optionally monosubstituted with R28 where R28 represents any value for R16 other than H;
A is selected from phenyl; naphthyl; a 5-10 membered monocyclic or bicyclic heteroaryl ring containing up to 5 heteroatoms where the heteroatoms are independently selected from O, N and S; or a xe2x80x94Sxe2x80x94Sxe2x80x94 dimer thereof when R2=H: or a N-oxide thereof; or an enantiomer, diastereoisomer, pharmaceutically acceptable salt, prodrug or solvate thereof together with a pharmaceutically acceptable diluent or carrier.
Preferably R1 is selected from H; xe2x80x94COxe2x80x94Oxe2x80x94(CH2)nPh optionally substituted on Ph as defined for R1=xe2x80x94C1-3alkylene-Ph and n=0-4; xe2x80x94COxe2x80x94Oxe2x80x94C2-4alkenyl; xe2x80x94COxe2x80x94C1-4alkyl; xe2x80x94C1-4alkylene-CONR4R5 where R4 and R5 are independently selected from H, C1-4alkyl.
Preferably R2 is selected from H and xe2x80x94COxe2x80x94C1-4alkyl.
Preferably L is selected from xe2x80x94CH2xe2x80x94NR18xe2x80x94; xe2x80x94CH2NR21xe2x80x94T.
Preferably A is selected from phenyl, naphthyl, pyridyl and thienyl.
Preferably combinations of R3 and p are selected from:
i) R3 is selected from a group of Formula II; xe2x80x94C1-4alkylR7; xe2x80x94Oxe2x80x94R7 and; R7; and p=1-3 with the proviso that one value of R3 is a group of Formula II;
ii) p=0 with the proviso that A is naphthyl and L is xe2x80x94CH2NR21xe2x80x94T;
iii) p=1 with the proviso that R1=a group of Formula II and A is naphthyl.
In another embodiment of the invention it is preferred that:
R1 is selected from H; xe2x80x94C1-4alkyl, xe2x80x94C1-3alkylene-Ph optionally mono or di-substituted on Ph with substituents selected from C1-4alkyl, halogen, OH, C1-4alkoxy, C1-4alkanoyl, C1-4alkanoyloxy, amino, C1-4alkylamino, di(C1-4alkyl)amino, C1-4alkanoylamino, thiol, C1-4alkylthio, nitro, cyano, carboxy, carbamoyl, C1-4alkoxycarbonyl, C1-4alkylsulfinyl, C1-4alkylsulfonyl, sulfonamido; xe2x80x94COxe2x80x94C1-4alkyl; xe2x80x94COxe2x80x94Oxe2x80x94C1-4alkyl; xe2x80x94COxe2x80x94Oxe2x80x94C2-4alkenyl; xe2x80x94COxe2x80x94Oxe2x80x94CH2-Ph optionally mono- or di-substituted on phenyl with substituents selected from C1-4alkyl, halogen, OH, C1-4alkoxy, C1-4alkanoyl, C1-4alkanoyloxy, amino, C1-4alkylamino, di(C1-4alkyl)amino, C1-4alkanoylamino, thiol, C1-4alkylthio, nitro, cyano, carboxy, carbamoyl, C1-4alkoxycarbonyl, C1-4alkylthiono, C1-4alkylsulfonyl, sulfonamido; xe2x80x94C1-4alkylene-CONR4R5 where R4 and R5 are independently selected from H, C1-4alkyl; xe2x80x94C1-4alkylene-COOR6 where R6 is selected from H, C1-4alkyl;
R2 is selected from H; xe2x80x94C1-4alkyl; xe2x80x94C1-3alkylene-Ph; xe2x80x94COC1-4alkyl; xe2x80x94COOC1-4alkyl:
R3 is selected from H; OH; CN; CF3; NO2; xe2x80x94C1-4 alkyl, xe2x80x94C1-4alkylene-R7 where R7 is selected from phenyl, naphthyl, a 5-10 membered monocyclic or bicyclic heteroaryl ring containing up to 3 heteroatoms selected from O, N and S; C1-4alkenyl; halogen; xe2x80x94(CH2)nCOOR8 where n=0-3 and R8 represents H, C1-4alkyl, C2-4alkenyl: xe2x80x94CONR9R10 where R9 and R10 independently represent H, C1-4alkyl, C2-4alkenyl, xe2x80x94Oxe2x80x94C1-4alkyl, xe2x80x94Oxe2x80x94C2-4alkenyl; xe2x80x94CON(R11)OR12 where R11 and R12 independently represent H, C1-4alkyl and C2-4alkenyl; a group of Formula II, xe2x80x94CONR13xe2x80x94CHR14xe2x80x94COOR17, where R13 is H or C1-4alkyl, R17 is H or C1-6alkyl, R14 is the side chain of a lipophilic amino acid with L or D configuration at the chiral alpha carbon in the corresponding free amino acid; C1-4alkyl monosubstituted on carbon with xe2x95x90Nxe2x80x94OH; xe2x80x94SOxe2x80x94C1-4alkyl; xe2x80x94SO2-C1-4alkyl; a group of Formula xe2x80x94Xxe2x80x94R15 where X is selected from CO, CH2, S, SO, SO2 and R15 is selected from C1-6alkyl, phenyl, naphthyl, a 5-10 membered monocyclic or bicyclic heteroaryl ring containing up to 3 heteroatoms selected from O, N and S;
p is 0-3 in which R3 values can be the same or different;
L is a linking moiety selected from the following groups written from left to right in Formula I; xe2x80x94COxe2x80x94NR16xe2x80x94 where R16 is selected from H, C1-4alkyl, C1-4alkylene-Z and Z is selected from xe2x80x94Oxe2x80x94C1-4alkyl, phenyl, naphthyl, a 5-10 membered monocyclic or bicyclic heteroaryl ring containing up to 3 heteroatoms selected from O, N and S; xe2x80x94CH2-NR18xe2x80x94 where R18 represents any value defined for R16; xe2x80x94CH2Sxe2x80x94; xe2x80x94CH2Oxe2x80x94; xe2x80x94CH2xe2x80x94CHR19xe2x80x94 where R19 represents any value defined for R16; xe2x80x94CHxe2x95x90CR20xe2x80x94 where R20 represents any value defined for R16; xe2x80x94CH2NR21xe2x80x94Txe2x80x94 where R21 represents any value defined for R16, T represents xe2x80x94(CH2)nxe2x80x94 where n is 1-4 and T is optionally monosubstituted with R22 where R22 represents any value for R16 other than H, and provided at least one of R21 and R22 is H; xe2x80x94CH2NR23xe2x80x94SO2xe2x80x94 where R23 represents any value defined for R16; xe2x80x94CH2-NR24xe2x80x94COxe2x80x94Txe2x80x94 where R24 represents any value defined for R16, T represents xe2x80x94(CH2)nxe2x80x94 where n is 0-4 and T is optionally monosubstituted with R29 where R29 represents any value for R16 other than H and provided at least one of R24 and R29 is H; xe2x80x94COxe2x80x94NR25xe2x80x94Txe2x80x94 where R25 represents any value defined for R16, T represents xe2x80x94(CH2)nxe2x80x94 where n is 1-4 and T is optionally monosubstituted with R26 where R26 represents any value for R16 other than H, and provided at least one of R24 and R25 is H; xe2x80x94CH2Sxe2x80x94Txe2x80x94 where T represents xe2x80x94(CH2)nxe2x80x94 where n is 1-4 and T is optionally monosubstituted with R27 where R27 represents any value for R16 other than H; xe2x80x94CH2Oxe2x80x94Txe2x80x94 where T represents xe2x80x94(CH2)nxe2x80x94 where n is 1-4 and T is optionally monosubstituted with R28 where R28 represents any value for R16 other than H;
A is selected from phenyl; naphthyl; a 5-10 membered monocyclic or bicyclic heteroaryl ring containing up to 3 or 5 heteroatoms in the case of monocyclic and bicyclic rings respectively where the heteroatoms are independently selected from O, N and S; or a xe2x80x94Sxe2x80x94Sxe2x80x94 dimer thereof when R2xe2x95x90H.
A preferred pharmaceutical composition is in the form of a tablet.
According to another aspect of the invention there is provided a compound of Formula I, III, IV or V for use as a medicament.
According to another aspect of the invention there is provided a compound of Formula I, III, IV or V for use in preparation of a medicament for treatment of a disease mediated through farnesylation of ras.
Many compounds of Formula I are a feature of this invention and in particular according to another aspect of the invention there is provided a compound of any of the following classes i), ii) or iii): 
wherein:
X1 is selected from H; C1-6alkyl: hydroxyC1-6alkyl, C1-6alkoxyC1-6alkyl C1-6alkylcarbonyl: hydroxyC1-6alkylcarbonyl: C1-6alkoxyC1-6alkylcarbonyl;
A is selected from phenyl, naphthyl or a 5-10 membered heterocyclic ring having up to 5 heteroatoms selected from O, N and S.
X2 is selected from H; phenyl: phenylC1-6alkyl: a 5-6 membered heteroaryl ring containing up to 3 heteroatoms selected from O, N and S optionally linked to A by C1-6alkyl; and X2 is optionally substituted on any ring as defined for phenyl in R1=xe2x80x94C1-3alkylene-Ph in claim 1;
X3 is selected from H; C1-6alkyl:
X4 is selected from C1-6alkylsulfanyl; C1-6alkylsulfinyl; C1-6alkylsulfonyl; carbamoyl; N-(C1-6alkyl)carbamoyl; N-(diC1-6alkyl)carbamoyl: and hydroxy or a C1-4alkyl ether thereof: 
wherein:
X5; is selected from xe2x80x94COxe2x80x94C1-4alkyl-Ph; xe2x80x94COxe2x80x94C1-6alkyl; xe2x80x94COxe2x80x94C1-6alkyl-heteroaryl where heteroaryl is a 5-10 membered heteroaryl ring containing up to 5 heteroatoms selected from O, N and S and Ph or heteroaryl are optionally substituted as defined for Ph in R1=xe2x80x94C1-3alkylene-Ph; C1-4alkyloxyC1-4alkyl;
A is naphthyl or a 10 membered heterocyclic ring having, up to 5 heteroatoms selected from O, N and S;
R3 and p are as defined in claim 1; 
wherein:
X6 has any value defined for X5 in ii) above;
X7 is Ph optionally substituted as defined for Ph in R1=xe2x80x94C1-3alkylene-Ph;
A is Ph or naphthyl or a 5-10 membered heterocyclic ring having up to 5 heteroatoms selected from O, N and S;
R3 and p are as defined above:
or a N-oxide, pharmaceutically acceptable salt, prodrug or solvate thereof.
Preferred values for compounds of class i) include,
X1 is selected from H and C1-6alkoxyC1-6alkyl;
X2 is selected from H; phenyl or phenylC1-6alkyl;
X4 is C1-6alkylsulfanyl;
A is selected from phenyl or naphthyl;
Other preferred values for X4 are xe2x80x94OMe and the lactone which can be formed when X4 is OH and X3 is H.
Preferred values for compounds of class ii) include p is 0.
Preferred values for compounds of class iii) include,
X7 is Ph;
A is Ph;
p is 0.
In another embodiment of the invention there is provided a compound of Formula I in which: R1 is selected from H or C1-4alkyl; R2 is selected from H, C1-4alkyl, xe2x80x94COC1-4alkyl; xe2x80x94C1-4alkylPh; L is selected from the following values as defined herein. CONR16, CH2S, CH2O, CH2CHR19, CHxe2x95x90CHR20, CH2NR24COT, CONR25T, CH2ST and CH2OT; and values for A, R3 and p are as defined herein, with the proviso that 2-(benzylcarbamoyl)4-sulfanylpyrollidine and 4-(acetylsulfonyl)-2(benzylcarbamoyl)-pyrrolidine are excluded. It is believed that the excluded compounds were disclosed as intermediates for beta-lactam antibiotic synthesis in Japanese patent application 60233076 (Sumitomo Chemical).
According to another aspect of the present invention there is provided any one of the following individual compounds or a pharmaceutically acceptable salt thereof:
(2S)-2-{2-Benzyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-benzoylamino}-4-methylsulfanylbutyric acid methyl ester;
(2S)-2-{2-Benzyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-benzoylamino}-4-methylsulfanylbutyric acid;
(2S)-2-({2-phenyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyric acid methyl ester;
(2S)-2-({2-phenyl-5-[([2S,4S]-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyric acid;
(2S)-2({3[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-naphthalene-1-carbonyl)}-amino)-4-methylsulfanylbutyric acid methyl ester;
(2S)-2-({3-[([2S,4S]-4-sulfanylpyolidin-2-ylmethyl)-amino]-naphthalene-1-carbonyl)}-amino)-4-methylsulfanylbutyric acid;
(2S)-2-({-3-phenyl-5[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyric acid methyl ester;
(2S)-2-({-3-phenyl-5[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyric acid;
(2S,4S)-2-[{N-(4-methoxybenzyl)-N-(naphthalen-1-ylmethyl)-amino}-methyl]-pyrrolidine-4-thiol;
N-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-pentanamide;
N-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-2-(pyridin-3-yl)-acetamide;
N-((2S,4S)-4-sulfanyl-pyrrolidin-2-ylmethyl)-3-methyl-N-(2-naphthalen-1-yl-ethyl)butyramide;
N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-N-(2-naphthalen-1-yl-ethyl)-2-pyridin-3-yl-acetamide;
(2S,4S)-2-{[(3-Methoxypropyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4-thiol;
N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-2-(4-methoxy-phenyl)-N-(2-naphthalen-2-yl-ethyl)-acetamide;
(2S,4S)-2-{[(2-(4-Methoxyphenyl)ethyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4-thiol;
N-(2,2-Diphenyl-ethyl)-N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3-methyl-butyramide;
N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-N-(2-naphthalen-2-yl-ethyl)-butyramide;
N-(2,2-Diphenyl-ethyl)-N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-butyramide;
(2S)-2-{3-[([2S,4S]-4-Sulfanyl-pyrrolidin-2-ylmethyl)-(3-methoxy-propyl)-amino]-benzoylamino}-4-methylsulfanyl-butyric acid;
N-([2S,4S]-4-Sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-N-(2-naphthalen-1-yl-ethyl)-butyramide:
(2S)-4-Carbamoyl-2-({2-phenyl-5-[([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-butyric acid; and
(2S)-4Carbamnoyl-2-({2-phenyl-5-[([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-butyric acid methyl ester.
According to another aspect of the invention there is provided a pharmaceutical composition comprising a compound as defined in any one Formulas III, IV or V or an individual compound listed above together with a pharmaceutically acceptable diluent or carrier.
According to another aspect of the invention there is provided a process for preparing compounds of classes i), ii) or iii) as defined above which comprises deprotecting a compound of Formula VI 
wherein X8 represents the right hand side of compound classes i), ii) or iii) as defined above. Pr1 is H or an amino protecting group. Pr2 is H or a thio protecting group and any functional groups in X8 are optionally protected with the proviso that there is at least one protecting group and optionally, if desired, converting the product thus obtained into a pharmaceutically acceptable salt thereof.
In an embodiment of the invention:
Examples of values for R1 include methyl; xe2x80x94CH2-Ph; xe2x80x94CH2-Ph substituted on Ph with nitro, especially 4-nitro; acetyl; BOC; allyloxycarbonyl; xe2x80x94COxe2x80x94Oxe2x80x94CH2-Ph substituted on Ph with nitro, especially 4-nitro; xe2x80x94CH2CONH2.
Examples of values for R2 include xe2x80x94COMe and xe2x80x94COOtertbutyl.
Examples of values for R3 include Cl; xe2x80x94COOH; xe2x80x94CONH; xe2x80x94SOMe and; xe2x80x94SO2Me.
When R3 represents xe2x80x94(CH2)xe2x80x94COOR8 a suitable value for n is 0.
Examples of lipophilic amino acids which contribute their side chain (denoted R14 within the definition of values for R3) include methionine, phenylglycine, phenylalanine. serine, leucine, isoleucine or valine. L configuration in the corresponding free amino acid is preferred. Examples of amino acid side chains are set out below. A preferred value for R14 is xe2x80x94CH2xe2x80x94CH2xe2x80x94Sxe2x80x94CH3. Further preferred values for R14 are xe2x80x94CH2xe2x80x94OMe and xe2x80x94CH2xe2x80x94CH2xe2x80x94OMe.
When R17is H to give a COOH group in Formula II, and R14 is xe2x80x94CH2xe2x80x94CH2xe2x80x94OH then a lactone can be formed where R17 and R14 together form part of a dihydrofuran-2-one heterocyclic ring. The same lactone can be formed for compounds of Formula III where X4 is OH and X3 is H.
A preferred value for p is 2.
When L is xe2x80x94CH2NR21xe2x80x94Txe2x80x94 a suitable value for n is 1. When L is xe2x80x94CH2xe2x80x94NR24xe2x80x94COxe2x80x94Txe2x80x94 a suitable value for n is 1. When L is xe2x80x94CH2xe2x80x94NR25xe2x80x94Txe2x80x94 a suitable value for n is 1. When L is xe2x80x94CH2xe2x80x94Sxe2x80x94Txe2x80x94 a suitable value for n is 1. When L is xe2x80x94CH2xe2x80x94Oxe2x80x94Txe2x80x94 a suitable value for n is 1. L is especially xe2x80x94CONHxe2x80x94, xe2x80x94CH2-NHxe2x80x94, xe2x80x94CH2NHSO2-, xe2x80x94CH2NHCOxe2x80x94.
Examples of values for A when A is heteroaryl are thienyl, pyridyl, quinolyl and quinoxalinyl.
Further preferred values are set out below.
For R1: 4-nitro-benzyloxycarbonyl; allyloxycarbonyl; carbamoylmethyl; acetyl; phenoxycarbonyl; H.
For R2: Acetylsulfanyl; H.
For R3: Methoxycarbonyl; N-methyl-N-methoxy-carbamoyl; nitro; allyloxycarbonyl; N-methyl-allyloxycarbamoyl; ethoxycarbonyl; 3,4-dichloro-benzyl-carbamoyl; hydroxy; carboxy; (2S),4-methylsulfanyl-butyric acid methyl ester-2yl-carbamoyl: (2S),4-methylsulfanyl-butyric acid-2yl-carbamoyl; phenoxy.
For p: 1-2, especially 2; a further preferred value is 0.
For L; xe2x80x94C(O)xe2x80x94NHxe2x80x94; xe2x80x94CH2C(O)xe2x80x94NHxe2x80x94; xe2x80x94CH2NHxe2x80x94C(O)xe2x80x94; xe2x80x94CH2-NHxe2x80x94SO2; especially xe2x80x94C(O)xe2x80x94NHxe2x80x94.
For A: phenyl; pyridyl, thienyl; naphthyl.
For R16 and R18-26: H, C1-4alkyl, especially H.
In another embodiment of the invention preferred values are set out below.
In compounds of Formula III: X1 is H or methoxyC1-4alkyl (especially H); X2 is H, phenyl or benzyl (especially benzyl); X3 is H or C1-4alkyl (especially H); X4 is C1-4alkylsulfanyl (especially methylsulfanyl); and A is phenyl. When A is a 6-membered aryl or heteroaryl ring then groups xe2x80x94NX1xe2x80x94 and the substituent comprising X4 are preferably in meta juxtaposition relative to each other; and X2, if present, is preferably positioned para relative to xe2x80x94NX1xe2x80x94. The chiral carbon to which xe2x80x94COOX3 is attached is preferably in S configuration. The chiral carbons at the 2 and 4 positions of the pyrrolidine ring are preferably in S configuration.
In compounds of Formula IV: X5 is xe2x80x94COxe2x80x94C1-4alkyl (especially xe2x80x94COxe2x80x94CH2xe2x80x94CHMe2) or xe2x80x94CH2xe2x80x94Phxe2x80x94Oxe2x80x94C1-4alkyl (especially xe2x80x94CH2xe2x80x94Phxe2x80x94OMe); heteroaryl is preferably pyridyl and a preferred aryl or heteroaryl substituent is xe2x80x94Oxe2x80x94C1-4alkyl (especially methoxy); and A is naphthyl. The chiral carbons at the 2 and 4 positions of the pyrrolidine ring are preferably in S configuration. The attachment point for A relative to xe2x80x94(CH2)1,2xe2x80x94 is preferably at the I position of napththalene and the equivalent position for heterocyclic values for A (regardless of ring numbering conventions for heterocycles). A preferred value for xe2x80x94(CH2)1,2xe2x80x94 is xe2x80x94(CH2)2xe2x80x94.
In compounds of Formula V: X6 is xe2x80x94COxe2x80x94C1-5alkyl (more preferably xe2x80x94COxe2x80x94CH2xe2x80x94CHMe2 or xe2x80x94COxe2x80x94CH2-t-butyl, especially xe2x80x94COxe2x80x94CH2xe2x80x94CHMe2) or xe2x80x94CH2xe2x80x94Phxe2x80x94Oxe2x80x94C1-4alkyl (especially xe2x80x94CH2xe2x80x94Phxe2x80x94OMe): heteroaryl is preferably pyridyl and a preferred aryl substitution is xe2x80x94Oxe2x80x94C1-4alkyl (especially methoxy): and A is phenyl or naphthyl (especially phenyl). The chiral carbons at the 2 and 4 positions of the pyrrolidine ring are preferably in S configuration. A preferred value for xe2x80x94(CH2)1,2xe2x80x94 is xe2x80x94(CH2)1xe2x80x94.
Suitable pairs of values for R3 when p=2 are; xe2x80x94COOMe, xe2x80x94CO.N(Me).OMe: NO2, xe2x80x94CO.N(Me).OMe; xe2x80x94COOMe, allyloxycarbonyl xe2x80x94CO.N(Me).OMe, allyloxycarbonyl; allyloxycarbonyl, xe2x80x94CO.N(Me).O.CH2CHxe2x95x90CH2: OH, COOH; xe2x80x94COOMe, COOMe: Ph, xe2x80x94CO.N-Methionine methyl ester: Ph, xe2x80x94CO.N-Methionine: benzyl, xe2x80x94CO.N-Methionine methyl ester; benzyl, xe2x80x94CO.N-Methionine; benzyl, xe2x80x94CO.N-Methionine isopropyl ester: Ph, xe2x80x94CO.Nxcex1-Glutamine methyl ester: Ph, xe2x80x94CO.Nxcex1-Glutamine.
Suitable values for Lxe2x95x90CHNR21T include CH2.N(CO.CH2.CHMe2).CH2.CH2; CH2.N(CH2CH2CH2OMe).CH2.CH2; CH2.N(CH2.pPh.OMe).CH2.CH2; CH2.N(CO.CH2.CHMe2).CH2; CH2N(CO.CH2.CH2.CH2.Me).CH2; CH2N(CO.CH2.CHMe.CH2Me).CH2; CH2N(CO.CH2.CH2.OMe)CH2; CH2N(CO.CH2.pyridin-3-yl).CH2; CH2N(4-methoxybenzyl)CH2; CH2N(CO.CH2.CHMe2)CH2.CH2.CH(Ph); CH2N(CO.CH3)CH2.CH2.CH(Ph); CH2N(CO.CH2.CHMe2)CH2; CH2N(CO.CH3)CH2; CH2N(CO.CH2.CHMe2)CH2.CH(Ph); CH2N(CO.CH2.CMe3)CH2.CH(Ph); CH2N(CO.CH2.pyridin-3-yl)CH2.CH(Ph); CH2N(CO.1-hydroxy-6-methoxy-pyridin-3-yl)CH2.CH(Ph); CH2N(CO.CH2CHMe2)CH2.CH2; CH2N(CO.CH2CMe3)CH2.CH2; CH2N(CO.CH2pyridin-3-yl)CH2.CH2; CH2N(CO.4-methoxybenzyl)CH2.CH2;
Suitable values for L=xe2x80x94CH2NR18xe2x80x94 include CH2NH: CH2NMe; CH2N(CO.CH2.CHMe2) and CH2N(CO.CH2.CH2.OMe).
Various forms of prodrug are well known in the art. For examples of such prodrug derivatives, see:
a) Design of Prodrug, edited by H. Bundgaard, (Elsevier, 1985) and Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al. (Academic Press, 1985);
b) A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 xe2x80x9cDesign and Application of Prodrugsxe2x80x9d, by H. Bundgaard p. 113-191 (1991);
c) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);
d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285 (1988); and
e) N. Kakeya, et al., Chem Pharm Bull. 32, 692 (1984).
Examples of pro-drugs include in vivo hydrolysable esters of a compound of the Formula I. An in vivo hydrolysable ester of a compound of the formula (I) containing
Some compounds within the scope of Formula I are known as intermediates in the human or animal body to produce the parent acid. Suitable pharmaceutically-acceptable esters for carboxy include C1-6alkoxymethyl esters for example methoxymethyl, C1-6alkanoyloxymethyl esters for example pivaloyloxymethyl, phthalidyl esters, C3-8cycloalkoxycarbonyloxyC1-6alkyl esters for example 1-cyclohexylcarbonyloxyethyl; 1,3-dioxolen-2-onylmethyl esters for example 5-methyl-1,3-dioxolen-2-onylmethyl; and C1-6alkoxycarbonyloxyethyl esters for example 1-methoxycarbonyloxyethyl and may be formed at any carboxy group in the compounds of this invention.
Particular substitutions on A for 6 membered rings are in the meta or para positions.
Some compounds within the scope of Formula I are known as intermediates in carbapenem side chain synthesis but it is believed that they have not been previously described in forms suitable as pharmaceutical compositions nor had any pharmaceutical activity associated with them per se. The reader is referred to the following publications in this regard and also in respect of synthetic details for compound preparation: Matsumura. Heterocycles (1995), 41, 147-59: European patent application EP 590885 (Zeneca; Betts et al.); European patent application EP 592167 (Zeneca; Siret); European patent application EP 562855 (Zeneca; Jung et al.); International patent application WO 92/17480 (Imperial Chemical Industries; Betts et al.); European patent application EP 508682 (Imperial Chemical Industries: Betts et al.): European Patent Application EP 280771 (Fujisawa Pharmaceutical, Murata et al.); and International patent application WO 92/17479 (Imperial Chemical Industries: Betts et al.).
In this specification the generic term xe2x80x9calkylxe2x80x9d includes both straight-chain and branched-chain alkyl groups. However references to individual alkyl groups such as xe2x80x9cpropylxe2x80x9d are specific for the straight-chain version only and references to individual branched-chain alkyl groups such as xe2x80x9cisopropylxe2x80x9d are specific for the branched-chain version only. An analogous convention applies to the other generic terms.
It is to be understood that, insofar as certain of the compounds of Formula I defined above may exist in optically active or racemic forms by virtue of one or more asymmetric carbon atoms, the invention includes in its definition any such optically active or racemic form which possesses the property of inhibiting FTPase. The synthesis of optically active forms may be carried out by standard techniques of organic chemistry well known in the art, for example by synthesis from optically active starting materials or by resolution of a racemic form. Similarly, inhibitory properties against FTPase may be evaluated using the standard laboratory techniques referred to hereinafter.
The term xe2x80x9chalogenxe2x80x9d refers to fluorine, chlorine, bromine and iodine. The term xe2x80x9ccarbamoylxe2x80x9d refers to xe2x80x94C(O)NH2. The term xe2x80x9cBOCxe2x80x9d refers to tert-butyl-Oxe2x80x94C(O)xe2x80x94. The ring systems in which both rings of the bicyclic system are aromatic.
Examples of C1-6alkyl include methyl, ethyl, propyl, isopropyl, sec-butyl, tert-butyl and pentyl; examples of C1-4alkyl include methyl, ethyl, propyl, isopropyl, sec-butyl and tert-butyl; examples of C1-3alkyl include methyl, ethyl, propyl and isopropyl; examples of xe2x80x94C1-3alkylenePh include benzyl, phenylethyl, phenylpropyl; examples of C1-4alkoxy (also called xe2x80x94Oxe2x80x94C1-4alkyl herein) include methoxy, ethoxy and propoxy; examples of C1-4alkanoyl include formyl, acetyl and propionyl; examples of C1-4alkanoyloxy include acetyloxy and propionyloxy; examples of C1-4alkylamino include methylamino ethylamino, propylamino, isopropylamino, sec-butylamino and tert-butylamino: examples of di-(C1-4alkyl)amino include di-methylamino, di-ethylamino and N-ethyl-N-methylamino; examples of C1-4alkanoylamino include acetamido and propionylamino; examples of C1-4alkoxycarbonyl include methoxycarbonyl, ethoxycarbonyl and propoxycarbonyl; examples of C1-4alkylsulfanyl include methylsulfanyl, ethylsulfanyl, propylsulfanyl, isopropylsufanyl, sec-butylsulfanyl and tert-butylsulfanyl; examples of C1-4alkylsulfinyl include methylsulfinyl, ethylsulfinyl, propylsulfinyl, isopropylsulfinyl, sec-butylsulfinyl and tert-butylsulfinyl: examples of C1-4alkylsulfonyl include methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, sec-butylsulfonyl and tert-butylsulfonyl examples of xe2x80x94COxe2x80x94C1-4alkyl include formyl, acetyl, propionyl, butyryl, and valeryl: examples of xe2x80x94COxe2x80x94Oxe2x80x94C1-4alkyl include ethyloxycarbonyl; propyloxycarbonyl and tert-butyloxycarbonyl (BOC); examples of xe2x80x94COxe2x80x94Oxe2x80x94C2-4alkenyl include allyloxycarbonyl and vinyloxycarbonyl; examples of xe2x80x94COxe2x80x94Oxe2x80x94(CH2)nPh where n=0-4 include phenyloxycarbonyl, benzyloxycarbonyl, phenylethyloxycarbonyl and phenylpropyloxycarbonyl; examples of xe2x80x94C1-4alkylene-CONR4R5 include carbamoylmethyl, carbamoylethyl, N-methylcarbamoylethyl, N-methyl-Nxe2x80x2-ethylcarbamoylethyl; examples of xe2x80x94C1-4alkylene-COOR6 include carboxymethyl, carboxyethyl, carboxypropyl, propionic acid methyl ester, acetic acid ethyl ester; examples of C2-4alkenyl include allyl and vinyl; examples of xe2x80x94Oxe2x80x94C2-4alkenyl include allyloxy and vinyloxy; examples of lipophilic amino acids include valine, leucine, isoleucine, methionine, phenylalanine, serine, threonine and tyrosine; examples of carbamoylC1-4alkyl include carbamoylmethyl, carbamoylethyl and carbamoylpropyl; examples of N-(monoC1-4alkyl)carbamoylC1-4alkyl include N-methyl-carbamoylmethyl and N-ethyl-carbamoylethyl; examples of N-(diC1-4alkyl)carbamoyl-C1-4alkyl include N,N-dimethylcarbamoylethyl and N-methyl-N-ethylcarbamoylethyl; examples of C1-4alkyl monosubstituted on carbon with xe2x95x90Nxe2x80x94OH include butyraldehyde oxime and propionaldehyde oxime: examples of hydroxyC1-6alkyl include hydroxymethyl, hydroxyethyl, hydroxypropyl, 2-hydroxypropyl, 2-(hydroxymethyl)propyl and hydroxypentyl; examples of C1-6alkoxyC1-6alkyl include methoxyethyl, ethoxyethyl and methoxybutyl; examples of C1-6alkylcarbonyl include methylcarbonyl, ethylcarbonyl propylcarbonyl, isopropylcarbonyl, sec-butylcarbonyl, tert-butylcarbonyl and pentylcarbonyl: examples of hydroxyC1alkylcarbonyl include hydroxyacetyl, hydroxypropionyl, hydroxybutyryl, 3-hydroxybutyryl and hydroxypentanoyl; examples of C1-6alkoxyC1-6alkylcarbonyl include methoxyacetyl, methoxypropionyl, ethoxybutyryl and butoxyacetyl; examples of phenylCtaylky include benzyl, phenylethyl and phenylpropyl: examples of xe2x80x94COxe2x80x94C1-4alkyl-Ph include phenylacetyl and phenylpropionyl; examples of xe2x80x94COxe2x80x94C1-4alkyl-heteroaryl include 2-(3-pyridyl)-acetyl and 2-(3-thienyl)-acetyl; examples of Nxe2x80x94(C1-6alkyl)carbamoyl include N-methyl-carbamoyl and N-ethyl-carbamoyl; examples of N-(diC1-6alkyl)carbamoyl include N,N-dimethylcarbamoyl and N-methyl-N-ethylcarbamoyl.
Examples of 5-10 membered monocyclic or bicyclic heteroaryl rings containing up to 5 heteroatoms selected from O, N and S include the following. Examples of 5- or 6-membered heteroaryl ring systems include imidazole, triazole, pyrazine, pyrimidine, pyridazine, pyridine, isoxazole, oxazole, isothiazole, thiazole and thiophene. A 9 or 10 membered bicyclic heteroaryl ring system is an aromatic bicyclic ring system comprising a 6-membered ring fused to either a 5 membered ring or another 6 membered ring. Examples of 5/6 and 6/6 bicyclic ring systems include benzofuran, benzimidazole, benzthiophene, benzthiazole, benzisothiazole, benzoxazole, benzisoxazole, pyridoimidazole, pyrimidoimidazole, quinoline, isoquinoline, quinoxaline, quinazoline, phthalazine, cinnoline and naphthyridine.
Preferably monocyclic heteroaryl rings contain up to 3 heteroatoms and bicyclic heteroaryl rings contain up to 5 heteroatoms. Preferred heteroatoms are N and S, especially N. In general, attachment of heterocyclic rings to other groups is via carbon atoms. Suitable values of heterocycles containing only N as the heteroatom are pyrrole, pyridine, indole, quinoline, isoquinoline, imidazole, pyrazine, pyrimidine, purine and pteridine.
Preferably any chiral carbon atoms at the 2 and 4 positions of the pyrrolidine ring in Formulas I and III-V are in S configuration.
Compounds of Formula I and III-V may form salts which are within the ambit of the invention. Pharmaceutically acceptable salts are preferred although other salts may be useful in, for example, isolating or purifying compounds.
When the compound contains a basic moiety it may form pharmaceutically acceptable salts with a variety of inorganic or organic acids, for example hydrochloric, hydrobromic, sulphuric, phosphoric, trifluoroacetic, citric or maleic acid. A suitable pharmaceutically-acceptable salt of the invention when the compound contains and acidic moiety is an alkali metal salt, for example a sodium or potassium salt an alkaline earth metal salt, for example a calcium or magnesium salt an ammonium salt or a salt with an organic base which affords a pharmaceutically-acceptable cation for example a salt with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine.
Solvates, for example hydrates, are also within the ambit of the invention and may be prepared by generally known methods.
According to another aspect of the present invention there is provided a compound of Formula I for use as a medicament.
According to another aspect of the present invention there is provided the use of a compound of Formula I in preparation of a medicament for treating ras mediated diseases, especially cancer.
According to another aspect of the present invention there is provided a method of treating ras mediated diseases, especially cancer, by administering an effective amount of a compound of Formula I to a mammal in need of such treatment.
According to a further feature of the invention there is provided a compound of Formula I, or a pharmaceutically-acceptable salt thereof, for use in a method of treatment of the human or animal body by therapy.
The invention also includes a method of treating a disease or medical condition mediated alone or in part by farnesylated ras which comprises administering to a mammal requiring such treatment an effective amount of an active ingredient as defined treatment of the human or animal body by therapy.
The invention also includes a method of treating a disease or medical condition mediated alone or in a part by farnesylated ras which comprises administering to a mammal requiring such treatment an effective amount of an active ingredient in the production of a new medicament for use in a farnesylated ras mediated disease or medical condition.
Specific cancers of interest include:
carcinoma, including that of the bladder, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid and skin;
hematopoietic tumors of lymphoid lineage, including acute lymphocytic leukemia, B-cell lymphoma and Burketts lymphoma;
hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias and promyelocytic leukemia;
tumors of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma; and
other tumors, including melanoma seminoma, tetratocarcinoma, neuroblastoma and glioma.
The compounds of Formula I are especially useful in treatment of tumors having a high incidence of ras mutation, such as colon, lung, and pancreatic tumors. By the administration of a composition having one (or a combination) of the compounds of this invention, development of tumors in a mammalian host is reduced.
Compounds of Formula I may also be useful in the treatment of diseases other than cancer that may be associated with signal transduction pathways operating through Ras, e.g., neuro-fibromatosis.
Compounds of Formula I may also be useful in the treatment of diseases associated with CAAX-containing proteins other than Ras (e.g., nuclear lamins and transducin) that are also post-translationally modified by the enzyme farnesyl protein transferase.
The compositions of the invention may be in a form suitable for oral use (for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (for example as creams, ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (for example as a finely divided powder or a liquid aerosol), for administration by insufflation (for example as a finely divided powder) or for parenteral administration (for example as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular or intramuscular dosing or as a suppository for rectal dosing).
The compositions of the invention may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art. Thus, compositions intended for oral use may contain, for example, one or more colouring, sweetening, flavouring and/or preservative agents.
Suitable pharmaceutically acceptable excipients for a tablet formulation include, for example, inert diluents such as lactose, sodium carbonate, calcium phosphate or calcium carbonate, granulating and disintegrating agents such as corn starch or algenic acid; binding agents such as starch; lubricating agents such as magnesium stearate, stearic acid or talc; preservative agents such as ethyl or propyl p-hydroxybenzoate, and anti-oxidants, such as ascorbic acid. Tablet formulations may be uncoated or coated either to modify their disintegration and the subsequent absorption of the active ingredient within the gastrointestinal tract, or to improve their stability and/or appearance, in either case, using conventional coating agents and procedures well known in the art.
Compositions for oral use may be in the form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate. calcium phosphate or kaolin. or as soft gelatin capsules in which the active ingredient is mixed with water or an oil such as peanut oil, liquid paraffin, or olive oil.
Aqueous suspensions generally contain the active ingredient in finely powdered form together with one or more suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia: dispersing or wetting agents such as lecithin or condensation products of an alkylene oxide with fatty acids (for example polyoxethylene stearate), or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives (such as ethyl or propyl g-hydroxybenzoate, anti-oxidants (such as ascorbic acid), colouring agents, flavouring agents, and/or sweetening agents (such as sucrose, saccharine or aspartame).
Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil (such as arachis oil, olive oil, sesame oil or coconut oil) or in a mineral oil (such as liquid paraffin). The oily suspensions may also contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set out above, and flavouring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.
Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water generally contain the active ingredient together with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients such as sweetening, flavouring and colouring agents, may also be present.
The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as for example liquid paraffin or a mixture of any of these. Suitable emulsifying agents may be, for example, naturally-occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soya bean, lecithin, an esters or partial esters derived from fatty acids and hexitol anhydrides (for example sorbitan monooleate) and condensation products of the said partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening, flavouring and preservative agents.
Syrups and elixirs may be formulated with sweetening agents such as glycerol, propylene glycol, sorbitol, aspartame or sucrose, and may also contain a demulcent, preservative, flavouring and/or colouring agent.
The pharmaceutical compositions may also be in the form of a sterile injectable aqueous or oily suspension, which may be formulated according to known procedures using one or more of the appropriate dispersing or wetting agents and suspending agents, which have been mentioned above. A sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example a solution in 1.3-butanediol.
Suppository formulations may be prepared by mixing the active ingredient with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Suitable excipients include, for example, cocoa butter and polyethylene glycols.
Topical formulations, such as creams, ointments, gels and aqueous or oily solutions or suspensions, may generally be obtained by formulating an active ingredient with a conventional, topically acceptable, vehicle or diluent using conventional procedure well known in the art.
Compositions for administration by insufflation may be in the form of a finely divided powder containing particles of average diameter of, for example, 30 xcexcor much less, the powder itself comprising either active ingredient alone or diluted with one or more physiologically acceptable carriers such as lactose. The powder for insufflation is then conveniently retained in a capsule containing, for example. 1 to 50mg of active ingredient for use with a turbo-inhaler device. such as is used for insufflation of the known agent sodium cromoglycate.
Compositions for administration by inhalation may be in the form of a conventional pressurised aerosol arranged to dispense the active ingredient either as an aerosol containing finely divided solid or liquid droplets. Conventional aerosol propellants such as volatile fluorinated hydrocarbons or hydrocarbons may be used and the aerosol device is conveniently arranged to dispense a metered quantity of active ingredient.
For further information on Formulation the reader is referred to Chapter 25.2 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch, Chairman of Editorial Board). Pergamon Press 1990.
The amount of active ingredient that is combined with one or more excipients to produce a single dosage form will necessarily vary depending upon the host treated and the particular route of administration. For example, a formulation intended for oral administration to humans will generally contain for example, from 0.5 mg to 2 g of active agent compounded with an appropriate and convenient amount of excipients which may vary from about 5 to about 98 percent by weight of the total composition. Dosage unit forms will generally contain about 1 mg to about 500 mg of an active ingredient. For further information on Routes of Administration and Dosage Regimes the reader is referred to Chapter 25.3 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.
The size of the dose for therapeutic or prophylactic purposes of a compound of the Formula I will naturally vary according to the nature and severity of the conditions, the age and sex of the animal or patient and the route of administration, according to well known principles of medicine. As mentioned above, compounds of the Formula I are useful in treating diseases or medical conditions which are due alone or in part to the effects of farnesylation of ras.
In using a compound of the Formula I for therapeutic or prophylactic purposes it will generally be administered so that a daily dose in the range, for example, 0.5 mg to 75 mg per kg body weight is received, given if required in divided doses. In general lower doses will be administered when a parenteral route is employed. Thus, for example, for intravenous administration, a dose in the range, for example, 0.5 mg to 30 mg per kg body weight will generally be used. Similarly, for administration by inhalation, a dose in the range, for example, 0.5 mg to 25 mg per kg body weight will be used. Oral administration is however preferred.
Compounds of this invention may be useful in combination with known anti-cancer and cytotoxic agents. If formulated as a fixed dose such combination products employ the compounds of this invention within the dosage range described herein and the other pharmaceutically active agent within its approved dosage range. Sequential use is contemplated when a combination formulation is inappropriate.
Although the compounds of the Formula I are primarily of value as therapeutic agents for use in warm-blooded animals (including man), they are also useful whenever it is required to inhibit the effects of activation of ras by farnesylation. Thus, they are useful as pharmacological standards for use in the development of new biological tests and in the search for new pharmacological agents.
According to another aspect of the present invention there is provided individual compounds produced as end products in the Examples set out below and salts thereof.
A compound of the invention, or a salt thereof, may be prepared by any process known to be applicable to the preparation of such compounds or structurally related compounds. Such processes are illustrated by the following representative schemes in which variable groups have any of the meanings defined for Formula I unless stated otherwise. Functional groups may be protected and deprotected using conventional methods. For examples of protecting groups such as amino and carboxylic acid protecting groups (as well as means of formation and eventual deprotection), see T. W. Greene and P. G. M. Wuts, xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d, Second Edition, John Wiley and Sons, New York, 1991. Note abbreviations used have been listed immediately before the Examples below.
Protecting groups may be removed by any convenient method as described in the literature or known to the skilled chemist as appropriate for the removal of the protecting group in question, such methods being chosen so as to effect removal of the protecting group with minimum disturbance of groups elsewhere in the molecule.
Specific examples of protecting groups are given below for the sake of convenience, in which xe2x80x9clowerxe2x80x9d signifies that the group to which it is applied preferably has 1-4 carbon atoms. It will be understood that these examples are not exhaustive. Where specific examples of methods for the removal of protecting groups are given below these are similarly not exhaustive. The use of protecting groups and methods of deprotection not specifically mentioned is of course within the scope of the invention.
A carboxy protecting group may be the residue of an ester-forming aliphatic or araliphatic alcohol or of an ester-forming silanol (the said alcohol or silanol preferably containing 1-20 carbon atoms).
Examples of carboxy protecting groups include straight or branched chain (1-12C)alkyl groups (e.g. isopropyl, t-butyl); lower alkoxy lower alkyl groups (e.g. methoxymethyl, ethoxymethyl, isobutoxymethyl; lower aliphatic acyloxy lower alkyl groups, (e.g. acetoxy,methyl, propionyloxymethyl, butyryloxymethyl, pivaloyloxymethyl); lower alkoxycarbonyloxy lower alkyl groups (e.g. 1-methoxycarbonylmethyl, 1-ethoxycarbonyloxyethyl); aryl lower alkyl groups (e.g. p-methoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, benzhydryl and phthalidyl); tri(lower alkyl)silyl groups (e.g. trimethylsilyl and t-butyldimethylsilyl); tri(lower alkyl)silyl lower alkyl groups (e.g. trimethylsilylethyl); and (2-6C)alkenyl groups (e.g. allyl and vinylethyl).
Methods particularly appropriate for the removal of carboxyl protecting groups include for example acid-metal- or enzymically-catalysed hydrolysis.
Examples of hvdroxy protecting groups include lower alkenyl groups (e.g. allyl); lower alkanoyl groups (eg. acetyl); lower alkoxycarbonyl groups (e.g. t-butoxycarbonyl); lower alkenyloxycarbonyl groups (e.g. allyloxycarbonyl); aryl lower alkoxycarbonyl groups (e.g. benzoyloxycarbonyl, p-methoxybenzyloxycarbonyl, o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl); tri lower alkyl/arylsilyl groups (e.g. trimethyisilyl, t-butyldimethylsilyl, tbutyldiphenylsilyl); aryl lower alkyl groups (e.g. benzyl) groups; and triaryl lower alkyl groups (e.g. triphenylmethyl).
Examples of amino protecting groups include formyl, aralkyl groups (e.g. benzyl and substituted benzyl. e.a. p-methoxybenzyl, nitrobenzyl and 2.4-dimethoxybenzyl, and triphenylmethyl); di-p-anisylmethyl and furylmethyl groups: lower alkoxycarbonyl (e.g. t-butoxycarbonyl): lower alkenyloxycarbonyl (e.g. allyloxycarbonyl); aryl lower alkoxycarbonyl groups (e.g. benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl: trialkylsilyl (e.g. trimethylsilyl and t-butyldimethylsilyl): alkylidene (e.g. methylidene): benzylidene and substituted benzylidene groups.
Methods appropriate for removal of hydroxy and amino protecting groups include for example, acid-base, metal- or enzymically-catalysed hydrolysis, or photolytically for groups such as o-nitrobenzyloxycarbonyl, or with fluoride ions for silyl groups.
Examples of protecting groups for amide groups include aralkoxymethyl (e.g benzyloxymethyl and substituted benzyloxymethyl): alkoxymethyl (e.g. methoxymethyl and trimethylsilylethoxymethyl); tri alkyl/arylsilyl (e.g. trimethylsilyl, t-butyldimethylsily, t-butyldiphenylsilyl); tri alkyl/arylsilyloxymethyl(e.g. t-butyldimethylsilyoxymethyl, t-butyldiphenyisilyloxymethyl); 4-alkoxyphenyl (e.g. 4-methoxyphenyl); 2,4-di(alkoxy)phenyl (e.g. 2.4-dimethoxyphenyl); 4-alkoxybenzyl (e.g. 4-methoxybenzyl); 2,4-di(alkoxy)benzyl (e.g. 2.4-di(methoxy)benzyl); and alk-1-enyl (e.g. allyl, but-1-enyl and substituted vinyl e.g. 2-phenylyinyl).
Aralkoxymethyl, groups may be introduced onto the amide group by reacting the latter group with the appropriate aralkoxymethyl chloride, and removed by catalytic hydrogenation. Alkoxymethyl, trialkyl/arylsilyl and trialkyl/silyloxymethyl groups may be introduced by reacting the amide with the appropriate chloride and removing with acid; or in the case of the silyl containing groups, fluoride ions. The alkoxyphenyl and alkoxybenzyl groups are conveniently introduced by arylation or alkylation with an appropriate halide and removed by oxidation with ceric ammonium nitrate. Finally alk-1-enyl groups may be introduced by reacting the amide with the appropriate aldehyde and removed with acid.
Compounds of Formula I in which L represents xe2x80x94COxe2x80x94NR16xe2x80x94 may be prepared by forming an amide bond between compounds 1 and 2 as outlined in Scheme 23 Compounds of Formula I in which L represents xe2x80x94COxe2x80x94NR25xe2x80x94Txe2x80x94 may be prepared by an analogous procedure. Suitable coupling conditions include the following.
i) Use of EEDQ at room temperature in an organic solvent (e.g. dichloromethane, methanol).
ii) Use of oxalyl chloride in an organic solvent (e.g. DMF. CH2Cl2) in the presence of an organic base (e.g. NMM. triethylamine, DMAP) at 0xc2x0 to room temperature for 0.5-16 h.
iii) Use of EDC/ HOBT in an organic solvent (e.g. DMF. CH2Cl2).
iv) Use of DCCI/ HOBT in an organic solvent (e.g. DMF. CH2Cl2) in the presence of an organic base (e.g. triethylamine).
v) Use of mixed anhydride reactions under standard conditions for example isopropylchloroformate in an organic solvent (e.g. DMF. DMA. dichloromethane) in the presence of an organic base (e.g. NMM. DMAP. triethylamine).
vi) Via an active ester under standard conditions e.g. pentafluorophenyl ester in an organic solvent (e.g. dichloromethane) in the presence of an organic base (e.g. triethylamine).
vii) Via an acid chloride under standard conditions e.g. using thionyl chloride and heat for about 150 min followed by an organic base (e.g. triethylamine) in the presence of an organic solvent (e.g. acetonitrile).
Compounds of Formula I in which L represents xe2x80x94CH2NR18xe2x80x94, xe2x80x94CH2Oxe2x80x94 or xe2x80x94CH2Sxe2x80x94 may be prepared as outlined in Scheme 24. LG represents a leaving group (e.g. mesyloxy, tosyloxy, halogen) and X represents S.O or NR18. Suitable coupling conditions include the following.
i) Use of an inorganic base (e.g. NaHCO3, NaH, K2CO3, butyllithium) in an organic solvent (e.g. THF, DMF, DMSO) and a temperature of about 70xc2x0 to 150xc2x0
ii) Use of an organic base (e.g. triethylam, DMAP) in an organic solvent (e.g. THF, dichloromethane, DMA, DMF) at a temperature range of room temperature xe2x88x92150xc2x0
iii) Use of an inorganic base (e.g. KOH, NaOH, K2CO,3) in an aqueous (e.g. water) and organic solvents (e.g. dichloromethane) in a 2 phase system, optionally in the presence of a phase transfer catalyst (e.g. tetrabutylammoniumbromide).
Compounds of Formula I in which L represents xe2x80x94CHxe2x95x90CR20xe2x80x94 may be prepared using a Wittig reaction as outlined in Scheme 25. Suitable reaction conditions include the following.
i) Use of a base (e.g. potassium carbonate, metal hydride, metal alkoxide) in the presence of an organic solvent (e.g. THF, toluene, DMSO) optionally in the presence of an aqueous solvent (2-phase system) and optionally in the presence of a catalyst complexing agent which solubilises alkali metal ions in non-polar solvents such as 1,4,7,10,13 -pentaoxacyclopentadecane ( also called 15-Crown-5) or 1,4,7,10,13,16 -hexaoxacyclooctadecane ( also called 18-Crown-6).
Compounds of Formula I in which L represents xe2x80x94CH2xe2x80x94NR18xe2x80x94 may be prepared as outlined in Scheme 26 by coupling aldehyde (2) with compound 4. Suitable coupling conditions include the following.
i) Use of a reducing agent (e.g. NaCNBH3, BH3, hydrogen plus catalyst, LiHBEt3xe2x80x94 di-isobutyl-aluminiumhydride, lithium aluminium hydride, sodium borohydride) in the presence of a suitable solvent e.g. ethanol and acetic acid.
Aldehyde (2) may be prepared by oxidation of the corresponding alcohol (1) under suitable conditions such as use of an oxidising agent (e.g. TPAP. NMMxe2x80x94O) in the presence of an organic solvent (e.g. acetonitrile, dichloromethane) at room temperature. Other suitable oxidising agents include chromium oxide, pyridinium chlorochromate, pyridinium dichromate, sodium dichromate and sodium hypochlorite.
Aldehyde (2) may also be prepared by reduction of the corresponding ester (1) under standard conditions using for example diisobutyl-aluminium hydride.
Compounds of Formula I in which L represents xe2x80x94CH2xe2x80x94NR21xe2x80x94Txe2x80x94. xe2x80x94CH2xe2x80x94Oxe2x80x94Txe2x80x94 or xe2x80x94CH2xe2x80x94Sxe2x80x94Txe2x80x94 may be prepared as outlined in Scheme 27 in which LG represents a leaving group (e.g. mesyloxy, tosyloxy, halogen) and X represent O, S or NR21. Suitable coupling conditions are as outlined above in relation to Scheme 24. Optionally the positions of LG and XH in compounds 1 and 2 in Scheme 27 can be reversed to give the same end product.
Compounds of Formula I in which L represents xe2x80x94CH2xe2x80x94NR23xe2x80x94SO2xe2x80x94 may be prepared as outlined in Scheme 28. Compounds 1 and 2 may be coupled under standard conditions such as the following.
i) Use of an organic base (e.g. di-isopropyl-ethylamine, triethylamine, 4-methyl-morpholine) in the presence of an organic solvent (e.g. dichloromethane) at a temperature range of 0xc2x0-40xc2x0
ii) Use of an inorganic base (e.g. potassium carbonate) in the presence of an organic solvent (e.g. DMF) at a temperature range of 0xc2x0-150xc2x0.
Compounds of Formula I in which L represents xe2x80x94CH2xe2x80x94NR24xe2x80x94COxe2x80x94Txe2x80x94 may be prepared as outlined in Scheme 29. Compounds 1 and 2 may be coupled under standard conditions such as described above for L xe2x95x90xe2x80x94COxe2x80x94NR16xe2x80x94.
Compounds of Formula I in which L represents xe2x80x94CH2xe2x80x94CHR 19xe2x80x94may be prepared as by reduction of compounds of the type set out as compound 3 in Scheme 25 but substituting R19 in lieu of R20. Reduction is carried out under standard conditions with standard reagents for example using hydrogenation in the presence of a catalyst such as palladium on charcoal at room temperature.
Biological activity was tested as follows. Farnesyl protein transferase (FPT) was partially purified from human placenta by ammonium sulphate fractionation followed by a single Q-Sepharose(copyright) (Pharmacia. Inc) anion exchange chromatography essentially as described by Ray and Lopez-Belmonte (Ray K P and Lopez-Belmonte J (1992) Biochemical Society Transations 20 494-497). The substrate for FPT was Kras (CVIM C-terminal sequence). The cDNA for oncogenic val 12 variant of human c-Ki-ras-24B was obtained from the plasmid pSW11-1(ATCC). This was then subcloned into the polylinker of a suitable expression vector e.g. plC 147. The Kras was obtained after expression in the E. coli strain, BL21. The expression and purification of c-KI-ras-24B and the vall2 variant in E. coli has also been reported by Lowe et al (Lowe P N et al. J. Biol. Chem. (1991) 266 1672-1678).
Incubations with enzyme contained 300 nM tritiated farnesyl pyrophosphate (DuPont/New England Nuclear), 120 nM ras-CVIM, 50 mM Tris HCl pH 8.0. 5 mM MgCl2, 10 xcexcM ZnCl2, 5 mM dithiotheitol and compounds were added at appropriate concentrations in DMSO (3% final concentration in test and vehicle control). Incubations were for 20 minutes at 37 xc2x0 and were stopped with acid ethanol as described by Pompliano et al. (Pompliano D L et al (1992) 31 3800-3807). Precipitated protein was then collected onto glass fibre filter mats (B) using a Tomtec(copyright) cell harvester and tritiated label was measured in a Wallac(copyright) 1204 Betaplate scintillation counter.
Although the pharmacological properties of the compounds of the Formula I vary with structural change as expected, in general compounds of the Formula I possess an IC50 in the above test in the range, for example, 0.01 to 200 xcexcM. Thus by way of example the compound 5{[(2S,4S)4-acetylsulfanyl-1-(4-nitro-benzyloxycarbonyl)-pyrrolidine-2-carbonyl]-amino}-3(N-methyl-methylcarbamoyl)-benzoic acid allyl ester (see Example 7) has an IC50 of approximately 0.5 xcexcM. No physiologically unacceptable toxicity was observed at the effective dose for compounds tested of the present invention.
The invention will now be illustrated in the following non-limiting Examples in which. unless otherwise stated:
(i) evaporations were carried out by rotary evaporation in vacuo and work-up procedures were carried out after removal of residual solids by filtration:
(ii) operations were carried out at room temperature, that is in the range 18-25xc2x0 C. and under an atmosphere of an inert gas such as argon:
(iii) column chromatography (by the flash procedure) and medium pressure liquid chromatography (MPLC) were performed on Merck Kieselgel silica (Art. 9385) or Merck Lichroprep RP-18 (Art. 9303) reversed-phase silica obtained from E. Merck. Darmstadt. Germany;
(iv) yields are given for illustration only and are not necessarily the maximum attainable;
(v) the end-products of the Formula I have satisfactory microanalyses and their structures were confirmed by nuclear magnetic resonance (NMR) and mass spectral techniques: chemical shift values were measured on the delta scale; the following abbreviations have been used: s, singlet; d. doublet; t or tr. triplet: m. multiplet: br. broad:
(vi) intermediates were not generally fully characterised and purity was assessed by thin layer chromatographic, infra-red (IR) or NMR analysis:
(vii) melting points are uncorrected and were determined using a Mettler SP62 automatic melting point apparatus or an oil-bath apparatus: melting points for the end-products of the Formula I were determined after crystallisation from a conventional organic solvent such as ethanol, methanol, acetone, ether or hexane, alone or in admixture: and
(viii) the following abbreviations have been used:
BOC tert-butoxycarbonyl
DCCI 1,3-dicyclohexylcarbodiimide
DMA N,N-dimethylacetamide
DMAP 4-dimethyl-aminopyridine
DMF N,N-dimethylformamide
DMSO dimethylsulfoxide
EDC 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide
EEDQ 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline
HOBT 1-hydroxybenzotriazole
NMM N-methylmorpholine
NMM-O 4-methylmorpholine-N-oxide
TFA trifluoroacetic acid
THF tetrahydrofuran
TMSI trimethylsilyliodide
TPAP tetrapropylammonium perruthenate
Note in the Schemes only those hydrogen atoms thought to assist clarity have been illustrated (ie not all hydrogen atoms have been illustrated).